Avaliação de anti-inflamatórios não esteroidais no tratamento da dor de ovinos submetidos à implantação de cânula ruminal e orquiectomia / Assessment of nonsteroidal anti-inflammatory drugs in treatment of pain in sheep submitted to ruminal cannulation and orchiectomy

O presente trabalho objetivou comparar o efeito do flunixin meglumine, cetoprofeno e meloxicam no tratamento da dor pós-operatória de ovinos submetidos à implantação de cânula ruminal e orquiectomia. Foram utilizados 32 ovinos, machos, pesando em média 35,5±3,5kg, distribuídos em três grupos: GFlu (flunixin meglumine 1,1mg/kg i.v.), GCet (cetoprofeno 3,0mg/kg i.v.) e GMel (meloxicam 0,5mg/kg i.v.). Exame clínico e coletas de sangue foram realizados no M0 (pré-avaliação), M1 (10 minutos após a pré-avaliação), M2 (início da sutura para fixação da cânula ruminal), M3 (logo após o término da cirurgia) e em duas, 12, 23, 25, 48 e 72 horas após a cirurgia (M2h, M12h, M23h, M25h, M48h e M72h), quando foram avaliados cortisol, glicose, proteína total, albumina, γ-glutamiltransferase (GGT), aspartato aminotransferase (AST), creatina quinase (CK), ureia, creatinina e hemograma. Nos M2h, M12h, M23h, M25h e M48h, foi realizada avaliação comportamental. O GFlu apresentou maior concentração de cortisol no M12h e no M48h e maior escore de dor na fístula e no testículo no M12h, quando comparado ao GMel. Os animais do GCet apresentaram menor interação com outros membros da baia no M23h. A ação analgésica do meloxicam foi maior em animais submetidos à implantação de cânula ruminal e orquiectomia, quando comparado ao flunixin meglumine e ao cetoprofeno.(AU)

This study aimed to compare the effect of flunixin meglumine, ketoprofen, and meloxicam in the treatment of postoperative pain in sheep submitted to ruminal cannulation and orchiectomy. 32 sheep were submitted to implantation of rumen cannula and orchiectomy, divided into three groups: GFlu (Flunixin meglumine 1,1mg/kg i.v.); GCet (Ketoprofen 3,0mg/kg i.v.); GMel (Meloxicam 0,5mg/kg i.v.). Clinical examination and blood samples were performed at M0 (pre-evaluation), M1 (10 minutes after pre-evaluation), M2 (beginning ruminal cannula), M3 (immediately post-surgery), and M2h, M12h, M23h, M25h, M48h and M72h (2h, 12h, 23h, 25h, 48h and 72 hours post-surgery) with the evaluation of cortisol, glucose, total protein, albumin, γ-glutamyl transferase (GGT), aspartate aminotransferase (AST), creatine kinase (CK), urea, creatinine and blood count. At M2h, M12h, M23h, M25h and M48h a behavioral evaluation was performed. The GFlu showed higher concentration of cortisol at M12h and M48h and greater pain score related with fistula and testis procedures at M12h when compared to GMel. Animals in the GCet group presented lower interaction with other animals in the same M23h paddock. The analgesia provided by Meloxicam was higher than flunixin meglumine and ketoprofen in animals submitted to placement of ruminal cannula and orchiectomy.(AU)

Hypertonic saline solution (NaCl 7.2%) enhances renal excretion of acids in cattle with acute ruminal lactic acidosis.

The present study aimed to evaluate the efficiency of hypertonic saline solution (HSS) as a novel treatment of acute ruminal lactic acidosis (ARLA) in cattle, focusing on urinary excretion of acids. Twelve cannulated steers were submitted to experimentally induced ARLA by adminis- tering sucrose into the rumen. Twenty hours later, the cattle were randomly divided into two equal groups. The first group was treated with 7.5% HSS (5 mL/kg) over 15 min, and isotonic saline solution (ISS; 20 mL/kg) for the subsequent 165 minutes. The control group was administered ISS instead of HSS. Rumen and urine samples were collected at different times during the experiment from the baseline to 64 h post-induction. The induction caused a medium-to-moderate ruminal acidosis, and a moderate degree of systemic acidosis and dehydration. Steers treated with HSS increased by 50% its glomerular filtration rate (1.61 mL/min) compared to ISS group (1.06 mL/ min; p⟨0.03). The overall volume of urine excreted by HSS group was higher than that in ISS group (1.62 L vs 0.7 L; p⟨0.02). This increase in total volume of urine provided by HSS favored a greater excretion of H+ ions in urine, which was 3.39-fold higher in HSS group (64.3*10-7 vs 18.9*10-7 Mol) as well as lactate (241.7 vs 181.8 mMol) and P urinary excretion (3.8 vs 1.1 mMol) that reduced the urine pH (5.3 vs 5.7). Only the HSS group decreased significantly blood total lactic acid concentration (20.3 %) throughout the treatment. A positive relationship was found between the excretion of urinary phosphorus and urinary pH (r2=0.562). The results showed that this novel treatment with HSS enhanced renal excretion of acids and may be recommended as an additional treatment for cattle with lactic acidosis.

Avaliação hemogasométrica, bioquímica e hematológica de ovinos suplementados com melão / Effects of sudden intake of melon on blood gas, biochemical and hematological parameters of sheep

O presente trabalho avaliou os efeitos da administração de duas diferentes quantidades de melão sobre variáveis hemogasométricas, bioquímicas e hematológicas de ovinos não adaptados. Foram utilizados 12 ovinos canulados, pesando 25kg de peso vivo, que nunca receberam ração concentrada. Os animais receberam dieta à base de feno (2,3% do peso vivo) e água à vontade. Os ovinos foram distribuídos aleatoriamente em dois grupos e receberam 25% ou 75% da matéria seca (MS) da dieta de melão triturado (G25% e G75%, respectivamente) diretamente no rúmen. Foram realizadas coletas de sangue e determinação do pH ruminal nos seguintes tempos: zero, 3, 6, 12, 18 e 24 horas após oferecimento do melão. Foi realizada análise hemogasométrica, do volume globular, determinação da concentração plasmática de lactato-L, glicose e osmolaridade sérica. No G25%, o pH sanguíneo variou entre 7,40 e 7,31, enquanto o G75% apresentou pH entre 7,38 e 7,26. Maiores concentrações de glicose plasmática foram detectadas no G75% no T3, T6 e T12 (P<0,05). Os ovinos que receberam 25% de melão mantiveram parâmetros sanguíneos dentro da normalidade, ao passo que, no G75%, os ovinos apresentaram discreta acidose metabólica sistêmica e hiperglicemia. A suplementação com 25% de melão pode ser uma alternativa segura na alimentação de ovinos.

This study evaluated the effects of two different amounts of melon on blood gas, biochemical and hematological variables of sheep not adapted. We used 12 cannulated sheep weighing 25 kg which never received concentrate. The animals received hay-based diet (2.3% of body weight) and water ad libitum. The sheep were randomly divided into two groups and received 25% or 75% of the dry matter (DM) of the diet of crushed melon (G25% and G75%, respectively) directly into the rumen. Blood collection and determination of ruminal pH were made at the following times: zero, three, six, 12, 18 and 24 hours after administration of the fruit. In whole blood was performed blood gas analysis and packed cell volume; in the plasma it was determined the concentrations of L-Lactate and glucose and in the serum the osmolarity. At G25% the blood pH ranged between 7.40 and 7.31, while G75% showed pH between 7.38 and 7.26. Higher concentrations of plasma glucose were detected in G75% after 3, 6 and 12 hours (P <0.05). Sheep receiving 25% of melon showed blood parameters within the normal range, while in the G75%, sheep had a mild systemic metabolic acidosis and hyperglycemia.

Behaviour of confined sheep fed with different concentrate sources / Comportamento ingestivo de ovinos confinados alimentados com diferentes fontes de concentrado

In this study we examined the effects of different feed concentrates on sheep behaviour. Our hypothesis was that citric pulp would stimulate rumination and be capable of replacing other concentrates traditionally used for feeding in confinement, to reduce the risk of urolithiasis. Ten adult Santa Inês sheep were distributed in a Latin square with five different diets, one control diet with 80 percent hay and 20 percent commercial feed and four diets containing 30 percent coast-cross hay and 70 percent of the following concentrates: pelleted citrus pulp, citrus pulp meal, cornmeal or wheat bran. After 21d of adaptation to each one of the five diets, the sheep were visually monitored for 24 h at 3 min intervals to record the time spent ruminating, time spent eating and time spent resting; the animals' positions (standing or lying down) were also noted. Daytime was considered to be from 06:00h to 18:00h. The data were evaluated using ANOVA, with Tukey post-hoc test or throughout Two-sample T test for circadian and position assessment. Citrus pulp diets resulted in time spent ruminating similar to the control diet (601, 590 and 669 min, respectively), but greater (P<0.05) than the cornmeal group (421min), which showed that citrus pulp generated effective rumination. The estimated saliva production in the control diet (26L) was greater than in the other groups, and was greater in the citrus pulp groups (24L/d) than cornmeal (21L/d). Feeding with cornmeal led to shorter time spent eating and time spent ruminating than all other diets. The sheep had higher time spent resting at night when fed concentrates (P<0.05). For all diets, about 90 percent of the time spent ruminating occurred with the animals lying down. Pelleted citrus pulp, citrus pulp meal and to a lesser degree wheat bran, led to adequate time spent ruminating. The use of citrus pulp can act as a preventive management measure to reduce the incidence of urolithiasis in sheep flocks...

No presente estudo, avaliaram-se os efeitos da alimentação de diferentes concentrados sobre o comportamento de ovinos. A hipótese é a de que a polpa cítrica estimularia a ruminação e reduziria o risco de ocorrência de urolitíase, podendo substituir outros concentrados. Dez ovinos adultos, mestiços da raça Santa Inês, foram distribuídos em um quadrado latino com cinco tratamentos, sendo quatro destes contendo dietas com 30 por cento de feno de capim coast-cross e 70 por cento dos seguintes concentrados: polpa cítrica peletizada, polpa cítrica farelada, fubá de milho e farelo de trigo, e uma dieta controle com 80 por cento de feno e 20 por cento de ração comercial peletizada. [...] A posição dos animais (em pé ou deitados) também foi observada. O período diurno foi considerado entre seis e 18 horas. Para comparação entre os tratamentos, os dados foram avaliados por meio de ANOVA e do teste de Tukey. Para a avaliação circadiana e entre as posições, foi utilizado o teste t de Student. Dietas com polpa cítrica promoveram tempo de ruminação semelhante aos do grupo de controle (601, 590 e 669 min, respectivamente), mas superior ao grupo alimentado com fubá de milho (421min). A produção de saliva estimada no grupo controle (26L/d) foi maior do que nos demais grupos, e os grupos com polpa cítrica tiveram maior produção de saliva do que o grupo com fubá de milho (21L/d). Ovinos em dietas ricas em concentrados descansam mais durante a noite. Em todas as dietas, cerca de 90 por cento da ruminação ocorreu com os animais deitados. A polpa cítrica peletizada e a farelada, e em menor grau o farelo de trigo, promoveram adequadamente a ruminação. Este concentrado pode ser utilizado como medida preventiva visando diminuir a incidência de urolitíase em rebanhos ovinos...

A long-term pulmonary rehabilitation program progressively improves exercise tolerance, quality of life and cardiovascular risk factors in patients with COPD.

BACKGROUND: Support and treatment options have been widely discussed in recent decades with the aim of improving morbidity, mortality and quality of life of chronic respiratory disease (COPD) patients. Although it is believed that longer pulmonary rehabilitation programs can provide better results, most of the evidence comes from short-term programs. AIM: To determine the effects of an outpatient pulmonary rehabilitation program on exercise tolerance, dyspnoea, hemodynamic variables and quality of life. DESIGN: Case series study. SETTING: Rehabilitation Centre. POPULATION AND METHODS: A convenience sample of COPD patients was enrolled in this study. The intervention consisted of a 96-wk exercise training program, including aerobic training, upper-limb exercises and inspiratory muscle training. Pulmonary function tests, blood biochemistry, six-minute walking distance test and health-related quality of life were recorded at baseline and after completion of the 6th, 12th, 18th, 24th months. RESULTS: Forty one consecutive COPD patients were recruited and thirty six completed the study. There was a significant improvement in hemodynamics, demonstrated by the gradual reduction in heart rate, blood pressure and MvO2 (double product) starting from the 12th month. Lipid profile showed a reduction of low density lipids and an increase of the high density lipids levels starting from the 6th month. Exercise tolerance, dyspnoea, respiratory muscle strength and quality of life also improved starting from the 6th month. CONCLUSION: A 24-month pulmonary rehabilitation program leads to a progressive improvement in quality of life, dyspnoea and exercise tolerance, and reduces cardiovascular risk factors in patients with chronic obstructive pulmonary disease. IMPACT: Our study suggests that long-term pulmonary rehabilitation programs can result in further improvements in the aforementioned cardiorespiratory variables.

Double-stranded RNA-activated protein kinase is a key modulator of insulin sensitivity in physiological conditions and in obesity in mice.

The molecular integration of nutrient- and pathogen-sensing pathways has become of great interest in understanding the mechanisms of insulin resistance in obesity. The double-stranded RNA-dependent protein kinase (PKR) is one candidate molecule that may provide cross talk between inflammatory and metabolic signaling. The present study was performed to determine, first, the role of PKR in modulating insulin action and glucose metabolism in physiological situations, and second, the role of PKR in insulin resistance in obese mice. We used Pkr(-/-) and Pkr(+/+) mice to investigate the role of PKR in modulating insulin sensitivity, glucose metabolism, and insulin signaling in liver, muscle, and adipose tissue in response to a high-fat diet. Our data show that in lean Pkr(-/-) mice, there is an improvement in insulin sensitivity, and in glucose tolerance, and a reduction in fasting blood glucose, probably related to a decrease in protein phosphatase 2A activity and a parallel increase in insulin-induced thymoma viral oncogene-1 (Akt) phosphorylation. PKR is activated in tissues of obese mice and can induce insulin resistance by directly binding to and inducing insulin receptor substrate (IRS)-1 serine307 phosphorylation or indirectly through modulation of c-Jun N-terminal kinase and inhibitor of κB kinase ß. Pkr(-/-) mice were protected from high-fat diet-induced insulin resistance and glucose intolerance and showed improved insulin signaling associated with a reduction in c-Jun N-terminal kinase and inhibitor of κB kinase ß phosphorylation in insulin-sensitive tissues. PKR may have a role in insulin sensitivity under normal physiological conditions, probably by modulating protein phosphatase 2A activity and serine-threonine kinase phosphorylation, and certainly, this kinase may represent a central mechanism for the integration of pathogen response and innate immunity with insulin action and metabolic pathways that are critical in obesity.

Migration of transjugular intrahepatic portosystemic shunt to the right atrium: complications in the intraoperative period of liver transplantation.

This article reports the case of a patient who underwent transjugular intrahepatic portosystemic shunt, which migrated to the right atrium. During liver transplantation, the extracardiac portion was sectioned and the portion adherent inside the atrium was managed expectantly.

Prevalence, treatment, and outcomes of the hepatic artery stenosis after liver transplantation.

INTRODUCTION: Hepatic artery stenosis (HAS) after liver transplantation can lead to altered hepatic function and/or thrombosis, there by increasing morbidity and mortality. The prevalence of HAS in the literatures varies from 4% to 11%. OBJECTIVE: We sought to describe the prevalence and treatment of hepatic artery stenosis. METHODS: We performed a descriptive retrospective analysis of 253 liver transplantations from March 1998 to May 2007, including patients with suspected HAS owing to increased hepatic enzymes, altered Doppler ultrasound (us) and hepatic biopsy. The confirmation of HAS was achieved through areriography. RESULTS: Nine patients were identified to have HAS, a 3.5% prevalence. Among the HAS patients, seven were male and two female. Their average age was 35.5 years (range, 65 to 53). The average time between the diagnosis and transplantation was 14.2 months (range, 9 to 68). The increase in hepatic enzymes among this group averaged: aspartate aminotransferase 131 U/L (range, 26 to 412) and alanine aminotransferase 192 U/L (range, 35 to 511). Doppler US showed alteration in the resistance level index. All patients underwent areriography; only one could not be treated owing to severe hepatic artery spasm, which also occurred during another attempt weeks after the first one. Among the eight patients, six were treated with stents and two with angioplastis. All treated patients displayed improvements in parameters. Four patients treated with stents required retreatment: two underwent angioplasty and two, a thrombolytic. One graft rethrombosed but evolved in compensated fashion with recanalization by collaterals. There has been no graft loss or mortality in this population. The average time of posttreatment follow-up was 31.28 (range, 9 to 68) months. CONCLUSION: The prevalence of HAS in our unit was within that reported in the literature. Treatment with a stent or angioplasty proved to be efficient to control this complication, considering that hepatic function recovered and that there was neither graft nor patient loss.

Differential gene expression assessed by cDNA microarray analysis in breast cancer tissue under tamoxifen treatment.

Our purpose was to identify tamoxifen (TAM) responsive genes after 30 days of TAM treatment in tumor tissues obtained from women with breast cancer using microarray expression analysis. In our study, we identified 12 candidates to be considered as tamoxifen-modulated genes. Among them, we selected two candidates the TEGT BI-1 (testis enhanced gene transcript Bax Inhibitor-1) and the CD63 gene in order to further confirm their differential expression under tamoxifen effects. We observed that both were down-regulated in tumor tissues of patients during TAM treatment. TEGT is able to inhibit the expression of Bax, which is known to promote apoptosis. On the other hand, CD63 encodes a cell membrane protein and it seems to be involved in mechanisms of platelet activation, cell adhesion and cell motility. We therefore hypothesize that TAM would be able to modulate tumor growth by down-regulating genes involved in mechanisms such as cell cycle control, tumor invasion and metastasis.

Complications following transjugular intrahepatic portosystemic shunt: a retrospective analysis.

Transjugular intrahepatic portosystemic shunt (TIPS) has been the therapeutic option for severe decompensation of chronic liver disease and as a bridge to liver transplantation. The aim of this study was to analyze the complications of this procedure. The records of 47 patients (39 men) of mean age 48 years underwent TIPS procedures from 1998 to 2003 were reviewed. Forty-one patients received 45 successful TIPS; it failed in six patients. Improvement was observed in 20 of 28 patients with upper gastrointestinal bleeding (71%); 9 of 11 with ascites (82%); and 5 of 8 with impaired renal function (62%). The Child-Pugh scores improved in 6 of the 47 patients (13%). Transplantation was performed in 11 patients (23%). The complications were: encephalopathy (49%); infection (19%); renal failure (17%); TIPS migration to the portal vein (4%) and to the right atrium (4%). Mortality was 32% (15/47) over 3 months. Eight patients developed active bleeding during TIPS installation requiring mechanical ventilation and intensive care, and died within the first week. Other causes of death were sepsis (n = 2), liver failure (n = 1), accidental puncture of the Glisson's capsule leading to intra-abdominal bleeding (n = 1) and refractory upper gastrointestinal bleeding (n = 3). The latter four patients had TIPS placement failure. In conclusion, TIPS produced clinical improvement among 51% of patients with complications in 49%. The main complications were encephalopathy (49%), infection (19%), and renal failure (17%). The 3-month mortality rate after TIPS placement was 32%.

PTX3 function as an opsonin for the dectin-1-dependent internalization of zymosan by macrophages.

Pentraxin 3 (PTX3) is a tumor necrosis factor and interleukin-1beta-stimulated gene that encodes a long PTX with proinflammatory activity. Here, we show that peritoneal macrophages derived from PTX3 transgenic (Tg) mice express higher levels of PTX3 mRNA than macrophages from wild-type (WT) mice, at basal level as well as upon stimulation with zymosan (Zy). Macrophages from Tg mice also showed improved opsonin-independent phagocytosis of Zy particles and the yeast form of the fungus Paracoccidioides brasiliensis. In the case of P. brasiliensis, an enhanced microbicidal activity accompanied by higher production of nitric oxide was also observed in macrophages from Tg mice. Using fluorescein-activated cell sorter analysis and reverse transcriptase-polymerase chain reaction, we demonstrated that basal level of Toll-like receptor-6 and Zy-induced dectin-1 expression was slightly but consistently higher in macrophages from Tg mice than in macrophages from WT mice. Recombinant (r)PTX3 protein binds to Zy particles as well as to yeast cells of P. brasiliensis and addition of rPTX3, to a culture of WT-derived macrophages containing Zy leads to an increase in the phagocytic index, which parallels that of Tg-derived macrophages, demonstrating the opsonin-like activity of PTX3. It is important that blockade of dectin-1 receptor inhibited the phagocytosis of Zy particles by WT and PTX3 Tg macrophages, pointing out the relevant role of dectin-1 as the main receptor involved in Zy uptake. Our results provide evidence for a role of PTX3 as an important component of the innate-immune response and as part of the host mechanisms that control fungal recognition and phagocytosis.

Identification, structural characterization, and tissue distribution of Tsg-5: a new TNF-stimulated gene.

Using DDRT-PCR, we compared the mRNA content of untreated and TNF-treated mouse embryonic fibroblasts (MEFs). Among differentially represented fragments, we identified and cloned a novel TNF-stimulated gene named Tsg-5. This gene, mapped to mouse chromosome 14, has three exons that can be alternatively spliced giving rise to two mRNA species, one spanning three exons and another that skips the second exon. Analysis of full-length Tsg-5 cDNA revealed a potential start codon within exon 2 encoding an ORF of 40 amino-acids. No homology with known mouse or human sequences, neither at the nucleotide nor at the amino-acid level could be found in public databases. In MEFs, Tsg-5 is induced by tumor necrosis factor-alpha (TNF) and IL-1 beta, albeit with distinct kinetics. TNF-induced Tsg-5 expression is NF-kappa B-dependent as it was inhibited by MG132, lactacystin, Bay 11-7083, and Bay 11-7085. Analysis of Tsg-5 expression in vivo revealed that the gene and its encoded polypeptide are constitutively expressed in the thymus and ovary, whereas, in LPS-treated mice, Tsg-5 mRNA can be detected in the spleen, lung, and brain. Our data suggest that Tsg-5 encodes a new, rare transcript, with a very tight regulation of expression and differential splicing.

Reduced susceptibility to collagen-induced arthritis in DBA/1J mice expressing the TSG-6 transgene.

OBJECTIVE: Expression of TSG-6 (tumor necrosis factor-stimulated gene 6) is induced by proinflammatory cytokines. This study was undertaken to examine the effects of local expression of TSG-6 in arthritic joints of TSG-6 transgenic mice, in the collagen-induced arthritis (CIA) model. METHODS: We generated transgenic mice that harbored the TSG-6 gene under the control of the T cell-specific lck promoter. Arthritis was induced by immunization with bovine type II collagen (CII), and its progression was monitored based on the incidence of arthritis, the arthritis index, and footpad swelling. Anti-CII antibodies and cytokine production were determined by enzyme-linked immunosorbent assay. Gene expression arrays were used to compare gene expression profiles of transgenic and control mice at various stages of CIA. RESULTS: TSG-6 was expressed in limbs of transgenic mice after immunization with CII, while its expression in nontransgenic animals was insignificant. The incidence of CIA was reduced in TSG-6 transgenic animals, its onset delayed, and all parameters of clinical arthritis significantly reduced. However, the immune response against CII was not significantly inhibited in TSG-6 transgenic mice. CONCLUSION: TSG-6 expression has been demonstrated in patients with rheumatoid and other forms of arthritis. Our data show that local expression of TSG-6 at sites of inflammation results in potent inhibition of inflammation and joint destruction in a model of autoimmune arthritis in mice. Therefore, it is likely that TSG-6 plays a similar modulatory role in human rheumatoid arthritis and related diseases and may have potential for the treatment of autoimmune arthritis in humans.

Quantitative and qualitative interferences of pentoxifillyne on hepatic Schistosoma mansoni granulomas: effects on extracellular matrix and eosinophil population.

Mast cells and eosinophils actively participate in tissue repair and are prominent components of Schistosoma mansoni granulomas. Since pentoxifillyne (PTX) is an immunomodulatory and antifibrotic substance, we aimed to characterize, by morphological techniques, the effect of this drug on fibrosis developed inside murine hepatic schistosomal granulomatous reaction, beyond the quantification of eosinophil and mast cell populations. The drug (1 mg/100 g animal weight) was administrated from 35 to 90 days post-infection, when the animals were killed. The intragranulomatous interstitial collagen network was analyzed by confocal laser scanning microscopy, the number of eosinophils and mast cells was quantified and the results were validated by t-student test. Treatment did not interfere on the granuloma evolution but caused a significant decrease in the total and involutive number of hepatic granulomas (p = 0.01 and 0.001, respectively), and in the intragranulomatous accumulation of eosinophils (p = 0.0001). Otherwise, the number of mast cells was not significantly altered (p = 0.9); however, it was positively correlated with the number of granulomatous structures (r = 0.955). In conclusion, PTX does not affect development and collagen deposition in S. mansoni murine granuloma, but decreases the intragranulomatous eosinophil accumulation possibly due to its immunomodulatory capability, interfering in cellular recruitment and/or differentiation.

Significant overexpression of oligophrenin-1 in colorectal tumors detected by cDNA microarray analysis.

The human oligophrenin-1 gene is ubiquitously expressed at low levels and expressed at high levels in the developing neuroepithelium of the neural tube. Mutations in this gene have been related to the X-linked mental retardation. Using cDNA microarrays, we found evidence that oligophrenin-1 is strongly up-regulated in colorectal tumors. Semiquantitative reverse transcriptase polymerase chain reaction confirmed this finding. Thus, a well-known nervous system-associated human gene transcript may also be an important colorectal tumor marker and potential therapeutic target.

TSG-14 transgenic mice have improved survival to endotoxemia and to CLP-induced sepsis.

Tumor necrosis factor-stimulated gene 14 (TSG-14)/PTX3 was identified originally as a TNF-alpha and IL-1beta-stimulated gene from normal, human foreskin fibroblasts and vascular endothelial cells, respectively. TSG-14 gene encodes a 42-kDa-secreted glycoprotein with a carboxy-terminal half that shares homology with the entire sequence of C-reactive protein (CRP) and serum amyloid P component (SAP), acute-phase proteins of the pentraxin family. Some experimental evidence suggests that TSG-14 plays a role in inflammation, yet its function and mechanism of action remain unclear. We have generated transgenic mice that overexpress the murine TSG-14 gene under the control of its own promoter. From eight transgenic founders, two lineages were derived and better characterized: Tg2 and Tg4, carrying two and four copies of the transgene, respectively. TSG-14 transgenic mice were found to be more resistant to the endotoxic shock induced by LPS and to the polymicrobial sepsis caused by cecal ligation and puncture (CLP). Moreover, macrophages derived from the transgenic mice produced higher amounts of nitric oxide in response to IFN-gamma, TNF-alpha, and LPS as compared with macrophages from wild-type animals, and the augmented response appears to be the consequence of a higher responsiveness of transgenic macrophages to IFN-gamma. The data shown here are the first in vivo evidence of the involvement of TSG-14 in the inflammatory process and suggest a role for TSG-14 in the defense against bacterial infections.

Sodium salicylate induces the expression of the immunophilin FKBP51 and biglycan genes and inhibits p34cdc2 mRNA both in vitro and in vivo.

One of the mechanisms proposed to explain the anti-inflammatory activity of sodium salicylate (NaSal) is based, at least in part, on its ability to inhibit nuclear factor-kappaB activation and inhibition of nuclear factor-kappaB-dependent gene expression. On the other hand, little is known about the ability of NaSal to activate gene expression. By differential display reverse transcription polymerase chain reaction, we identified several genes that are modulated upon treatment of mouse fibroblasts with NaSal. From the various cDNA fragments recovered from autoradiograms, we found that NaSal can increase the levels of mRNA for biglycan, the mouse homologue of the human eIF-3 p47 unit, and immunophilin FKBP51. NaSal-induced expression of these genes was time- and dose-dependent. Moreover, FKBP51 gene expression was augmented in vivo, in mice treated orally or intraperitoneally with NaSal. We also found that treating cells with NaSal can inhibit the expression of the p34(cdc2) kinase. The impact this inhibition on cell cycle was evaluated by measuring the content of DNA during the cell cycle. Treatment of cells with NaSal led to a G(2)/M arrest. By investigating the signaling events that regulate the expression of these genes and their biological activities, we can contribute to the understanding of the mechanism of NaSal.

Shotgun sequencing of the human transcriptome with ORF expressed sequence tags.

Theoretical considerations predict that amplification of expressed gene transcripts by reverse transcription-PCR using arbitrarily chosen primers will result in the preferential amplification of the central portion of the transcript. Systematic, high-throughput sequencing of such products would result in an expressed sequence tag (EST) database consisting of central, generally coding regions of expressed genes. Such a database would add significant value to existing public EST databases, which consist mostly of sequences derived from the extremities of cDNAs, and facilitate the construction of contigs of transcript sequences. We tested our predictions, creating a database of 10,000 sequences from human breast tumors. The data confirmed the central distribution of the sequences, the significant normalization of the sequence population, the frequent extension of contigs composed of existing human ESTs, and the identification of a series of potentially important homologues of known genes. This approach should make a significant contribution to the early identification of important human genes, the deciphering of the draft human genome sequence currently being compiled, and the shotgun sequencing of the human transcriptome.

Differential regulation of TSG-14 expression in murine fibroblasts and peritoneal macrophages.

Tumor necrosis factor (TNF)-stimulated gene 14 (TSG-14, also termed PTX3) encodes a secreted glycoprotein whose carboxy-terminal half shares sequence similarity with the pentraxin family of acute phase proteins (C-reactive protein and serum amyloid P component). We compared TSG-14 mRNA expression in cultures of murine BALB/c 3T3 fibroblasts and thioglycollate-elicited peritoneal macrophages. TNF and interleukin-1 (IL-1) potently induced TSG-14 expression in 3T3 fibroblasts but not in peritoneal macrophages. Lipopolysaccharide (LPS) elicited TSG-14 expression in both cell types, but induction in 3T3 cells and macrophages showed several distinct characteristics. Whereas in 3T3 fibroblasts TSG-14 mRNA was rapidly up-regulated by LPS, expression in macrophages was substantially delayed. Furthermore, cycloheximide greatly reduced LPS-induced TSG-14 mRNA up-regulation in macrophages but not in 3T3 cells. Finally, interferon-gamma (IFN-gamma; but not IFN-alpha/beta) inhibited LPS-induced TSG-14 expression in macrophages and not in 3T3 fibroblasts. The antioxidant pyrrolidine dithiocarbamate inhibited LPS-induced nuclear factor-kappaB (NF-kappaB) activation and TSG-14 expression in macrophages. In contrast, IFN-gamma did not inhibit NF-kappaB function as measured by IkappaB-alpha and IkappaB-beta degradation, IkappaB-alpha resynthesis, or electrophoretic mobility shift analysis. Inhibition of LPS-induced TSG-14 mRNA expression by IFN-gamma in macrophages was also observed in the presence of cycloheximide and in cells from STAT1 null mice, suggesting that IFN-gamma inhibits TSG-14 expression through an unconventional mechanism.

Schistosoma mansoni egg-induced hepatic granulomas in mice deficient for the interferon-gamma receptor have altered populations of macrophages, lymphocytes and connective tissue cells.

Systemic production and mobilization of inflammatory cells and formation of hepatic periovular granulomas were studied in Schistosoma mansoni-infected mice with deficient interferon gamma (IFN-gamma) receptor (IFN-gammaR(o/o)). The impaired IFN-gamma signaling did not cause a significant modification of the overall kinetics of inflammatory cells, but mutant mice developed smaller hepatic periovular granulomas with a two-fold reduction in all the cell lineages. In granulomas of normal mice, the fully differentiated macrophages were progressively predominant, whilst in IFN-gammaR(o/o) mice, the granulomas contained a higher percentage of immature and proliferating monocytes. Granulomas of IFN-gammaR(o/o) mice had an enhanced and accelerated fibrotic reaction, corresponding to an increased content of proliferative and activated connective tissue cells. Simultaneously, their granulomas had an increased ratio of T over B cells, with an increase in CD8(+) and a reduction in CD4(+) T cells. The functional IFN-gamma receptor was not required for initial recruitment of monocytes and lymphocytes into granulomas, but it was necessary for the maturation of macrophages, upregulation of major histocompatibility class 2 (MHC-II) expression and consequent stimulation of lymphocyte subpopulations depending upon the MHC-II-mediated antigen presentation.